Influenza virus vaccines and uses thereof
The present disclosure provides influenza hemagglutinin stem domain polypeptides comprising (a) an influenza hemagglutinin HA1 domain that comprises an HA1 N-terminal stem segment, covalently linked by a linking sequence of 0-50 amino acid residues to an HA1 C-terminal stem segment, and (b) an influenza hemagglutinin HA2 domain, wherein on or more amino acids in the HA2 domain have been mutated. Also provided are nucleic acids encoding the polypeptides, compositions comprising the polypeptides and/or nucleic acid molecules, as well as methods of their use, in particular in the detection, prevention and/or treatment of influenza.
1. A method for producing an influenza hemagglutinin stem domain polypeptide, the method comprising:
removing the cleavage site between HA1 and HA2 of an influenza HA0 peptide from an influenza virus comprising HA of the H1 subtype by mutating the C-terminal amino acid of HA1 into an amino acid other that arginine (R) or lysine (K);
removing the peptide of the globular head domain comprising at least amino acids 53 to 320 from the HA0 sequence;
introducing one or more mutations in the peptide connecting the C-terminal residue of helix A to the N-terminal residue of helix CD comprising the amino acid sequence MNTQX 1 TAX 7 GKEX 3 N(H/K)X 4 E(K/R) (SEQ ID NO: 190), wherein one or more of the amino acids X 1 , X 2 , X 3 and X 4 has been changed into an amino acid selected from the group consisting of serine (S), threonine (T), asparagine (N), glutamine (Q), R, histidine (H), K, aspartic acid (D), glutamic acid (E), and glycine (G); and
introducing a disulfide bridge in the HA stem domain polypeptide between the amino acids at position 324 and 436,
so as to produce the influenza hemagglutinin stem domain polypeptide,
wherein the numbering is based on the numbering of amino acids of the H1N1 influenza strain A/Brisbane/59/2007 (SEQ ID NO: 1).
2. The method according to claim 1 , wherein the C-terminal amino acid residue of the HA1 C-terminal stem segment is mutated into Q, S, T, N, D or E.
3. The method according to claim 1 , wherein the C-terminal amino acid residue of the HA1 C-terminal stem segment is mutated into Q.
4. The method according to claim 1 , wherein
X 1 has been changed into S,
X 2 has been changed into T, Q, or G,
X 3 has been changed into S, and
X 4 has been changed into S.